The purpose of this study was to investigate the relationship between organizational culture and affective well-being at work and to investigate the moderating role of person-organization (P–O) fit in this relationship. Data were obtained from an experimental design through a questionnaire using scenarios with 127 police officers. No interaction effect between organizational culture and P–O fit has been verified. Nevertheless, a principal effect was obtained regarding P–O fit (i.e., a higher P–O fit seems to promote higher levels of affective well-being at work, especially comfort and enthusiasm). The findings encourage future research on the relationship between P–O fit and affective well-being at work in police organizations.